LOCUS Exported 5550 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION N-terminal tagging with mTFP1 using LEU2 as a selection marker in S. cerevisiae.. ACCESSION . VERSION . KEYWORDS POM43-mTFP1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5550) AUTHORS Higuchi-Sanabria R, Garcia EJ, Tomoiaga D, Munteanu EL, Feinstein P, Pon LA TITLE Characterization of Fluorescent Proteins for Three- and Four-Color Live-Cell Imaging in S. cerevisiae. JOURNAL PLoS One. 2016 Jan 4;11(1):e0146120. doi: 10.1371/journal.pone.0146120. eCollection 2016. PUBMED 26727004 REFERENCE 2 (bases 1 to 5550) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5550 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind complement(53..86) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind complement(2375..2408) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS 2447..3151 /codon_start=1 /product="monomeric teal (cyan) fluorescent protein (Ai et al., 2006)" /label=mTFP1 /note="mammalian codon-optimized" /translation="VSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDGT NTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTFE DKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLKG DVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESAV ARNSTDGMDELYK" primer_bind complement(3187..3206) /label=T7 /note="T7 promoter, forward primer" promoter complement(3188..3206) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3293..3310) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3464..4052) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3544..3563) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4223..5083) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 4846..4865 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5084..5188) /gene="bla" /label=AmpR promoter primer_bind 5256..5274 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(5312..5334) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 5434..5453 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 5534..2 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 5534..1 /label=SP6 /note="SP6 promoter, forward primer" ORIGIN 1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acaaccctta atataacttc 61 gtataatgta tgctatacga agttattagg tctagagatc cgcaggctaa ccggaacctg 121 tattatttag tttatgctac gttaaataaa gacctttcgt tcacataact gaatgtgtaa 181 tggccttgag atttcaagca taccaagttg gtggagacgg ggtcgttaca aaagactctt 241 taagccaaga tttccttgac agccttggcg atagcatcgc caacctcagt ggtagagtta 301 gaaccaccaa ggtcaccggt tctgacacct gcatccaaga catttctaac agcttcttca 361 agagccctac cttcttcaac caaatccaag gataacttca acatcatagc tgcagataag 421 atggtagcaa ttgggttaac cttgtttgct ggtaaatctg gggcagaacc atgacatggt 481 tcgtacaaac cgaatgcctt gttagtgtca ggtagggaag ctagagatgc agaaggtaat 541 aaacccaaag aacctggaat aacagaggct tcatcggaga taatatcacc aaacatgttg 601 ttggtaataa caacaccgtt tagcttagtt ggtgatttaa ccaaaatcat agcagcagag 661 tcgatcaatt ggtgctgaac agttaattgt gggaactcag tcttgatggt ttcttcaaca 721 gtctttctcc acaatctgga agaggcaagc acgttagcct tgtcaagaga ccagattggt 781 aatggtgggt tttgttgcaa tgccaagaaa gcagccattc ttgtaattct ttgaacttca 841 ggaacactgt atttctcaga gtcccaagca actccgtcac cttcatcttc ttttctttca 901 ccaaagtaga taccaccaac caattctcta acgacgacga aatcggtacc ctttgcatat 961 tcaggcttca aaggagaaag atctagtaaa gaatcagaag caaagttaca tggtctcaag 1021 ttggcgtata gacccaattc ctttctgatc ttcaatagac cttgttctgg tctaactgcg 1081 cccgtacccc attttggacc accaacagca cctagtaaga cagcatcggc tttcttagag 1141 gcttctagag cttcatctgg taaaggagtg ccagtggcat cgatggcagc acccccgatc 1201 aagtgatgtt ggaaattgaa cttaatttct ggacggactt cagcaatggc attcaagacc 1261 ttaatagctt cgtcagtaac ttctttaccg acgtgatcac ccggtaggac aacgatattc 1321 ttagacatgg tataatctgt gtagtgtggg atacttttta cttcttcaaa taggtatcaa 1381 caataaaaat aaatcgaagc aaatgtagga atgcgttaaa gcagatgtac ttacttagag 1441 tacatagata tatttatata attcaatata taaaagtata tgaagcatct gatgttgaac 1501 ctgtcatgac tctaaactgg aatgggtagt tatgcttctg aaggttttcc gttagaaaat 1561 gagtgcaggg atcaggcccg gaaccggttt tagcctgaaa ggaaaagaaa aaaatgcggt 1621 gattactagc acgtgactgc gctgaattgg aatccaccac aagtacagcg aatggcaaga 1681 gggaagggga atatattgtg cctggcggta gtctgctata tcaaatgtta aacataacta 1741 aatgcagtag tgaatagagt aatgtatttc tacatatgtg ggtagggcca atgggagccc 1801 gatgtctatt gtacagatat tttcctttta tagttaatca attcaacttg gaaatcattg 1861 agtcgatatc tacttggata actggagccg tggcagcgtt tgaaatttga cgagtcttct 1921 tgttttgttc gcgcattttc agtttattca aacgggagat ggcaccgaat ccactgactt 1981 ctctgttcaa tttgaatggt aataggaaga aagtacattt cgagaaattt tcgcccaacg 2041 tcgaaatctc atggatgata tcctcgatac aaatgacacc ttcatcacct aatttctctt 2101 caacgatatt gttgtcattc aagattatct cttttggttc tttatcttct ggcctttgcc 2161 acattattct agatctcttt tggattaaag aacggataga tgcaagtgaa ggttgtccga 2221 taacaatata tggtgataca agtttcaaca atgggtacac gtccttggta agtttgatga 2281 aaacaccagt gttcaactca acaagtctca aaagagataa cactttgtaa gcctttgctg 2341 ggatcttcac agcgagctct cgagaaccct taatataact tcgtataatg tatgctatac 2401 gaagttatta ggtgatatca tgcgtacgct gcaggtcgac ggatccgtga gcaagggcga 2461 ggagaccaca atgggcgtaa tcaagcccga catgaagatc aagctgaaga tggagggcaa 2521 cgtgaatggc cacgccttcg tgatcgaggg cgagggcgag ggcaagccct acgacggcac 2581 caacaccatc aacctggagg tgaaggaggg agcccccctg cccttctcct acgacattct 2641 gaccaccgcg ttcgcctacg gcaacagggc cttcaccaag taccccgacg acatccccaa 2701 ctacttcaag cagtccttcc ccgagggcta ctcttgggag cgcaccatga ccttcgagga 2761 caagggcatc gtgaaggtga agtccgacat ctccatggag gaggactcct tcatctacga 2821 gatacacctc aagggcgaga acttcccccc caacggcccc gtgatgcaga agaagaccac 2881 cggctgggac gcctccaccg agaggatgta cgtgcgcgac ggcgtgctga agggcgacgt 2941 caagcacaag ctgctgctgg agggcggcgg ccaccaccgc gttgacttca agaccatcta 3001 cagggccaag aaggcggtga agctgcccga ctatcacttt gtggaccacc gcatcgagat 3061 cctgaaccac gacaaggact acaacaaggt gaccgtttac gagagcgccg tggcccgcaa 3121 ctccaccgac ggcatggacg agctgtacaa gactagtggc ctatgcggcc gcggatctgc 3181 cggtctccct atagtgagtc gtattaattt cgataagcca ggttaacctg cattaatgaa 3241 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 3301 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 3361 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 3421 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 3481 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 3541 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 3601 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat 3661 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 3721 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 3781 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 3841 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 3901 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 3961 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 4021 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 4081 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 4141 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 4201 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 4261 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 4321 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 4381 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 4441 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 4501 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 4561 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 4621 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 4681 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 4741 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 4801 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 4861 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 4921 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 4981 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 5041 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 5101 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 5161 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct 5221 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc 5281 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg 5341 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg 5401 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag 5461 tgcaccatat ggacatattg tcgttagaac gcggctacaa ttaatacata accttatgta 5521 tcatacacat acgatttagg tgacactata //